Clostridium difficile is an anaerobic, spore- forming and gram-positive bacteria. The major virulence factors of this bacteria are A and B toxin that encoded by tcdA and tcdB genes. In this study, stool speciments was collected from diarrheal children (n=112) . In orther to detection of A and B toxin and tpigenes,were assessed by multiplex PCR method. In this study C.difficile were identified in 22/32% of patients that toxigenic strain was 68% . The pupose of this study, was simuntaneous detection of tcdA and tcdB genes, in order to identification of toxigenic C.difficile strains in diarrheal children by multiplex PCR